Jiang Zengxin, Jiang Chang, Jin Lixia, Chen Zixian, Feng Zhenzhou, Jiang Xiaoxing, Cao Yuanwu
Department of Orthopedic Surgery, Zhongshan Hospital, Fudan University, Shanghai, China.
J Orthop Res. 2022 Oct;40(10):2350-2361. doi: 10.1002/jor.25264. Epub 2022 Jan 19.
Diabetes mellitus contributes to intervertebral disc degeneration. Nucleus pulposus cell senescence plays an important role in intervertebral disc degeneration. However, the effects of hyperglycemia on human nucleus pulposus cells and the underlying process remains poorly understood. In the current study, we evaluated the effects of high glucose levels on human nucleus pulposus cell senescence in vitro and the effects of hyperglycemia on rat nucleus pulposus aging in vivo. Human nucleus pulposus cells were cultured in high-glucose medium (200 mM glucose) for 48 h. Senescence-associated β-galactosidase staining, western blot analysis, and enzyme-linked immunosorbent assays were performed to evaluate human nucleus pulposus cell senescence. Flow cytometry and enzyme-linked immunosorbent assays were used to evaluate reactive oxygen species and advanced glycation end-product levels. Transcriptome sequencing followed by bioinformatics analysis was used to understand the abnormal biological processes of nucleus pulposus cells cultured in high-glucose medium. Diabetes mellitus rat models were established and histopathological and immunohistochemical analysis was conducted to examine nucleus pulposus tissue senescence in vivo. Exposure to a high glucose concentration promoted human nucleus pulposus cell senescence and increased the senescence-related secretion phenotype in human nucleus pulposus cells in vitro and in rat nucleus pulposus tissue in vivo. Bioinformatics analysis showed that hub genes were involved in nucleus pulposus cell cycle activities and cell senescence. The results suggest that appropriate blood glucose control may be key to preventing intervertebral disc degeneration in diabetic patients.
糖尿病会导致椎间盘退变。髓核细胞衰老在椎间盘退变中起重要作用。然而，高血糖对人髓核细胞的影响及其潜在机制仍知之甚少。在本研究中，我们评估了高糖水平对体外培养的人髓核细胞衰老的影响以及高血糖对体内大鼠髓核衰老的影响。将人髓核细胞在高糖培养基（200 mM葡萄糖）中培养48小时。进行衰老相关β-半乳糖苷酶染色、蛋白质印迹分析和酶联免疫吸附测定以评估人髓核细胞衰老。使用流式细胞术和酶联免疫吸附测定来评估活性氧和晚期糖基化终产物水平。通过转录组测序并进行生物信息学分析，以了解在高糖培养基中培养的髓核细胞的异常生物学过程。建立糖尿病大鼠模型，并进行组织病理学和免疫组织化学分析以检查体内髓核组织衰老情况。暴露于高糖浓度会促进体外培养的人髓核细胞衰老，并增加体外培养的人髓核细胞和体内大鼠髓核组织中与衰老相关的分泌表型。生物信息学分析表明，枢纽基因参与髓核细胞周期活动和细胞衰老。结果表明，适当控制血糖可能是预防糖尿病患者椎间盘退变的关键。
